Project Summary/Abstract: Immune Monitoring & Flow Cytometry (IMFC) Immune Monitoring and Flow Cytometry were separate CCSG-funded shared resources that were combined in July 2010 into a single shared resource, the Immune Monitoring & Flow Cytometry (IMFC) Shared Resource. For ease of reference, the unit publicizes its services to clients as DartLab. Flow Cytometry is located at both the Lebanon and Hanover sites, and Immune Monitoring is located within Norris Cotton Cancer Center's (NCCC) Lebanon facilities. Equipment offered by Flow Cytometry includes a single FACSAria I cell sorter operated by a dedicated flow cytometrist at the Lebanon site, in addition to a 10-color Gallios, 7-color MACSQuant, 4-color FACSCalibur and 3-color FACScan. There is an 8-color MACSQuant VYB and a 6-color FACSCanto at the Hanover site. Equipment offered by Immune Monitoring includes a Zeiss enzyme-linked immunoSpot assay (ELISPOT) reader, two multiplex cytokine analyzers: a Bio-Plex Array Reader and a Sector Imager (MSD); two automated magnetic bead cell separators: an autoMACS and a Robosep; and an 8-color MACSQuant 10. Services, used by both clinicians and basic scientists, include peripheral blood mononuclear cell isolation and cryopreservation, plasma/serum isolation and cryopreservation, multiplex cytokine analysis, lymphoproliferation, ELISPOT, up to 10-color phenotyping, including multimers and intracellular staining; and customized immunoassays. Flow Cytometry has allowed multiple NCCC PIs to sort cell subsets, including Ernst (CM), Dmitrovsky (CM/MT), Noelle (ICI), and Wang (ICI). The use of multicolor flow cytometry has allowed Stan (CM), Dmitrovsky (CM/MT), Turk (ICI), Ernstoff (ICI), and Mullins (ICI) to subset tumor-infiltrating cells, especially regulatory T cells. Multiplex cytokine analysis has been used by multiple NCCC PIs, including Karagas (CE), Ernstoff (ICI), Sentman (ICI), Turk (ICI), Mullins (ICI), and Stan (CM). Immune Monitoring has customized assays for Lansigan (MT), Kinlaw (MT), and Noelle (ICI). The IMFC is directed by Dr Jacqueline Channon Smith, who holds a Ph.D. in innate immunity, overseeing two experienced laboratory managers, one of whom is the dedicated cell sorter cytometrist; a research associate who holds a Ph.D. in immunology, and specialists who are cross-trained in flow cytometry and immunoassay techniques. The IMFC is used by a total of 65 principal investigators across all six NCCC programs: Cancer Control (1), Cancer Epidemiology (3), Cancer Mechanisms (17), Molecular Therapeutics (20), Cancer Imaging and Radiobiology (5), and Immunology and Cancer Immunotherapy (19).